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Nitrogen inputs influence vegetative metabolism in maize engineered with a seed-specific carotenoid pathway

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Abstract

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Metabolomic profiling of a maize line engineered with an endosperm-specific carotenogenic pathway revealed unexpected metabolic readjustments of primary metabolism in leaves and roots.

Abstract

High-carotenoid (HC) maize was engineered to accumulate high levels of carotenoids in the endosperm. The metabolic interventions influenced the flux through non-target pathways in tissues that were not affected by the targeted intervention. HC maize at the vegetative stage also showed a reduced susceptibility to insect feeding. It is unknown, however, whether the metabolic history of the embryo has any impact on the metabolite composition in vegetative tissues. We, therefore, compared HC maize and its isogenic counterpart (M37W) to test the hypothesis that boosting the carotenoid content in the endosperm triggers compensatory effects in core metabolism in vegetative tissues. Specifically, we investigated whether the metabolite composition of leaves and roots at the V6 stage differs between HC and M37W, and whether N inputs further alter the core metabolism of HC compared to M37W. We found an increase in the abundance of organic acids from the tricarboxylic acid (TCA) cycle in HC even under restricted N conditions. In contrast, low levels of carotenoids and chlorophyll were measured regardless of N levels. Sugars were also significantly depleted in HC under low N. We propose a model explaining the observed genotype-dependent and input-dependent effects, in which organic acids derived from the TCA cycle accumulate during vegetative growth and contribute to the increased demand for pyruvate and/or acetyl-CoA in the endosperm and embryo. This response may in part reflect the transgenerational priming of vegetative tissues in the embryo induced by the increased demand for metabolic precursors during seed development in the previous generation.

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Acknowledgements

We thank Dr. Roxana Savin for helping with the design of nitrogen treatments, as well as Jose Perez, Derry Alvarez, Pedro Cerdá and Dr. Teresa Capell for helping with the preparation of soil substrates. Special thanks to Jaume Capell for all the assistance provided in the greenhouse. P.C. acknowledges support from the ERA-NET SUSCROP PROSTRIG project funded by the Spanish Ministry of Science, Innovation and Universities (project number PCI2019-103382). P.D.F. and L.P.-F. are grateful to DEFRA for funding the PROSTRIG project through the ERA-NET SUSCROP scheme (project number CH0217). P.S.G.-C. was supported through an Agrotecnio postdoctoral fellowship.

Funding

P.C. acknowledges support from the ERA-NET SUSCROP PROSTRIG project funded by the Spanish Ministry of Science, Innovation and Universities (Project Number PCI2019-103382). P.D.F. and L.P.-F. are grateful to DEFRA for funding the PROSTRIG project through the ERA-NET SUSCROP scheme (project number CH0217). P.S.G.-C. was supported through an Agrotecnio postdoctoral fellowship.

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Authors and Affiliations

  1. Department of Plant Production and Forestry Sciences, University of Lleida-Agrotecnio Center, Lleida, Spain

    Patricia S. Girón-Calva & Paul Christou

  2. Department of Biological Sciences, Royal Holloway, University London, Egham, Surrey, UK

    Laura Pérez-Fons & Paul D. Fraser

  3. Institute of Molecular Bioscience, J. W. Goethe University, Frankfurt am Main, Germany

    Gerhard Sandmann

  4. ICREA, Catalan Institute for Research and Advanced Studies, Barcelona, Spain

    Paul Christou

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  1. Patricia S. Girón-Calva
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  2. Laura Pérez-Fons
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  3. Gerhard Sandmann
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  4. Paul D. Fraser
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  5. Paul Christou
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Contributions

PC conceived the original idea; PSG-C designed and performed the experiments in the greenhouse; PSG-C prepared samples for GCMS and UPLC analysis; LP-F conducted the GCMS and UPLC analysis. PSG-C analyzed the data and wrote the article; LP-F supervised the data analysis; PDF and GS assisted in data interpretation; PC supervised the writing and edited the article. PDF and PC agree to serve as the authors responsible for contact and ensure communication.

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Correspondence to Paul D. Fraser or Paul Christou.

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Communicated by Stefan Schillberg.

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Supplementary Information

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299_2021_2689_MOESM1_ESM.docx

Supplementary file 1 Online Resource 1. Metabolite profile and changes in metabolite concentrations in M37W and HC leaves under low-nitrogen (N−) and normal (N +) input regimens. Online Resource 2. Metabolite profile and changes in metabolite concentrations in M37W and HC roots under low-nitrogen (N−) and normal (N +) input regimens. Online Resource 3. Plant material and N treatments used in this study. (DOCX 959 KB).

299_2021_2689_MOESM2_ESM.xlsx

Supplementary file 2 Online Resource 4. Data sets generated during and/or analyzed during the current study. (XLSX 36 KB).

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Girón-Calva, P.S., Pérez-Fons, L., Sandmann, G. et al. Nitrogen inputs influence vegetative metabolism in maize engineered with a seed-specific carotenoid pathway. Plant Cell Rep 40, 899–911 (2021). https://doi.org/10.1007/s00299-021-02689-2

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